Abstract
Elevated levels of proinflammatory cytokines associated with infection and inflammation can modulate CYP450 enzymes leading to potential disease-drug interactions and altered small molecule drug disposition. We established a human derived hepatocyte-Kupffer cell (Hep:KC) co-culture model to assess the indirect cytokine impact on hepatocytes through stimulation of Kupffer cell-mediated cytokine release and compared this model to hepatocytes alone. Characterization of Hep:KC co-cultures showed an inflammation response following treatment with LPS and IL-6 (indicated by secretion of various cytokines). Additionally, IL-6 exposure up-regulated acute phase proteins (CRP, AAG and SAA2) and down-regulated CYP3A4. Compared to hepatocytes alone, Hep:KC co-cultures showed enhanced IL-1β-mediated effects but less impact from both IL-2 and IL-23. Hep:KC co-cultures treated with IL-1β exhibited a higher release of proinflammatory cytokines, an increased up-regulation of acute phase proteins and a larger extent of metabolic enzyme and transporter suppression. IC50 values for IL-1β-mediated CYP3A4 suppression were lower in Hep:KC co-cultures (98.0-144 pg/mL) compared to hepatocytes alone (IC50 >5000 pg/mL). CYP suppression was preventable by blocking IL-1β interaction with IL-1R1 using an antagonist cytokine or an anti-IL-1β antibody. Unlike IL-1β, IL-6-mediated effects were comparable between hepatocyte monocultures and Hep:KC co-cultures. IL-2 and IL-23 caused a negligible inflammation response and a minimal inhibition of CYP3A4. In both hepatocyte monocultures and Hep:KC co-cultures, IL-2RB and IL-23R were undetectable while IL-6R and IL-1R1 were higher in Hep:KC co-cultures. In summary, compared to hepatocytes monocultures, the Hep:KC co-culture system is a more robust in vitro model for studying the impact of proinflammatory cytokines on metabolic enzymes.
- cell models
- cytochrome P450
- cytokines/chemokines
- drug-drug interactions
- hepatocytes
- inflammation
- Kupffer cells
- The American Society for Pharmacology and Experimental Therapeutics